Objective To observe the efficacy of stage treatment with acupuncture plus intermediate frequency for Bell's palsy and its effect on serum NO content. Method By adopting a randomized controlled design, eighty eligible patients with Bell's palsy were randomized into a treatment group and a control group. The two groups both received acupuncture treatment in acute stage (within 1 week after the onset); in the remission stage (since 1 week after the onset), electroacupuncture apparatus was added for the control group, and intermediate frequency was given to the treatment group in addition to the intervention for the control group. The House-Brackmann (H-B) facial nerve grading system and serum NO content were observed before and after the treatment. Result The H-B facial nerve grading was improved after the intervention in both groups; the total effective rate was 95.0% in the treatment group, significantly higher than 82.5% in the control group (P<0.05); the serum content of NO was significantly improved after the treatment in both groups (P<0.05), while the increase in the treatment group was more significant than that in the control group (P<0.05). Conclusion In treating Bell's palsy, stage treatment with Acupuncture plus intermediate frequency can effectively restore facial nerve function, significantly enhance clinical efficacy and shorten treatment duration. This disease is closely related to the content of serum NO, and the benign regulation on serum NO content is possibly the mechanism of acupuncture in treating facial paralysis.

Objective To investigate the relationship between the mechanism of ischemic postconditioning-induced activation of nuclear factor-E2 related factor 2 (Nrf2)-antioxidant response element (ARE) signaling pathway during myocardial ischemia-reperfusion (I/R) and reactive oxygen species (ROS).Methods Healthy male Sprague-Dawley rats, aged 16-20 weeks, weighing 250-300 g, were heparinized and anesthetized with intraperitoneal 1％ pentobarbital sodium 40 mg/kg.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution.Thirty-two isolated rat hearts were randomly divided into 4 groups (n=8 each) using a random number table: control group (group C) , group I/R,ischemic postconditioning group (group IPO) , and N-(2-mercaptopropionyl)-glycine (a ROS scavenger) + IPO group (group M + IPO).After 20 min of equilibration, group C was continuously perfused with K-H solution for 100 min, and the isolated hearts received the drugs via the perfusion system in the other groups.Group I/R was perfused with cardioplegic solution 4 ℃ St.Thomas, and then was subjected to 40 min of ischemia at 32 ℃ followed by 60 min of reperfusion.In group IPO, ischemic postconditioning was induced by 6 cycles of 10 s reperfusion followed by 10 s limb ischemia starting from the onset of reperfusion, and the hearts were then perfused for 58 min.In group M + IPO, the hearts were perfused with K-H solution containing N-(2-mercaptopropionyl)-glycine 2 m mol/L for 3 min starting from the onset of reperfusion,underwent 2 min of ischemic postconditioning, and then was perfused for 55 min.Heart rate (HR), left ventricular developed pressure (LVDP), left ventricular end-diastolic pressure (LVEDP),and positive maximal pressure of left ventricular increase (+dp/dtmax) were recorded at the end of equilibration and of reperfusion.At 5 min of reperfusion and the end of reperfusion, myocardial specimens were obtained from the left ventricle for determination of ROS content by enzyme-linked immunosorbent assay.At the end of reperfusion, myocardial specimens were obtained from the left ventricle for examination of the ultrastructure of myocardial cells and for determination of Nrf2, heme oxygenase-1 (HO-1) , quinone oxidoreductase 1 (NQO1), and superoxide dismutase 1 (SOD1) mRNA and protein expression (by using Western blot and real-time polymerase chain reaction).The damage to myocardial mitochondria was assessed using Flameng scoring.Results Compared with group C, HR, +dp/dtmax and LVDP were significantly decreased, and LVEDP was increased at the end of reperfusion in I/R and M+IPO groups, HR and LVDP were decreased, LVEDP was increased, and no significant changes were found in +dp/dtmax at the end of reperfusion in IPO group, Flameng score was increased in I/R, IPO and M+IPO groups , the ROS content was increased at the end of reperfusion in I/R, IPO and M+IPO groups, and Nrf2, HO-1,NQO1 and SOD1 mRNA and protein expression was down-regulated at the end of reperfusion in I/R, IPO and M+IPO groups.Compared with group I/R, HR, +dp/dtmax and LVDP were significantly increased, and LVEDP and ROS content were decreased at the end of reperfusion, Nrf2, HO-1, NQO1 and SOD1 mRNA and protein expression was up-regulated at the end of reperfusion in IPO and M+IPO groups, Flameng score was decreased in IPO group, there was no significant change in Flameng score in M+IPO group.Compared with group IPO, HR, +dp/dtmax and LVDP were significantly decreased, LVEDP and ROS content were increased at the end of reperfusion, Flameng score was increased, and Nrf2, HO-1, NQO1 and SOD1 mRNA and protein expression was down-regulated in M+IPO group.Conclusion Ischemic postconditioning can regulate ROS level and activate Nrf2-ARE signaling pathway, thus attenuating myocardial I/R injury in rats.

Objective To observe the activation mechanism of Nrf2-ARE pathway in protective effect of ischemia and pinacidil postconditioning on isolated rat hearts.Methods The hearts of adult male Sprague Dawley rats were established ischemia-reperfusion injury model,and devided into six groups(n =8,each group),i.e.Normal group(Group N),ischemiareperfusion group (Group Con,I/R),ischemic postconditioning group (Group IPO),pinacidil postconditioning group (Group P50),N-(2-mercaptopropionyl)-glycine(MPG,2mmol/L) + IPO group(Group M + IPO),MPG + P50 group(Group M + P50).Rat hearts were perfused with Krebs-Henseleit(K-H) buffer for 20 minutes for equilibration.Subsequently,Group N was perfused with K-H buffer for 100 minutes after equilibration,Group Con was perfused with 4℃ ST.Thomas solution to stop the heart beating after equilibration,then the hearts were underwent 40 minutes global ischemia under 32℃,and followed by the K-H solution for 60 minutes.Group IPO after global ischemia period,the hearts were subjected to six 10-seconds cycles of ischemia/reperfusion at the beginning of reperfusion,then were reperfused for 58 minutes.Group P50 after global ischemia,rat hearts were perfused with K-H buffer containing pinacidil(50.μmol/L) for 2 minutes before reperfusion.Group M + IPO after global ischemia,the hearts were subjected to perfuse with K-H buffer containing MPG(2 mmol/L) for 3 minutes,and then underwent six 10-seconds cycles of ischemia/reperfusion before reperfusion.Group M + P50 after global ischemia,the hearts were perfused with K-H buffer containing MPG(2 mmol/L) for 3 minutes,and then subjected to perfuse with K-H buffer containing pinacidil(50 μmol/L) for 2 minutes before reperfusion.Cardiac function indexes(such as HR,LVDP,LVEDP,and the Max dp/dt) at the end point of equilibration and repeffusion were observed and recorded.The ultrastructure of myocardial tissue was observed by electron microscopy and the mitochondrial Flameng score was calculated.RT-PCR and western-blot were applied to detect the gene transcription and protein expression of HO-1,NQO1,SOD1,and Nrf2 in left ventricular myocardial tissue after reperfusion.Results The HR,LVDP and + dp/dtmax at the end of reperfusion:the cardiac function indexes are lower among each group compared with group N,group 1PO and group P50 are better than group Con (P ＜ 0.05).Compared with group IPO,there is no significant difference in group group P50,but group M + IPO is obviously decreased(P ＜ 0.05).Compared with group P50,group M + P50 index is decreased significantly(P ＜ 0.05).The LVEDP at the end of reperfusion is lower than that among each group as compared with group Con,which is significantly increased in group Con (P ＜ 0.05).Compared with group IPO,there is no significant difference in group P50,but group M + IPO is significantly increased(P ＜ 0.05).Compared with group P50,the group M + P50 is obviously decreased(P ＜ 0.05).The ultrastructure of myocardial tissue in group N is mostly normal,group Con presence serious damage.The ultrastructure damage of myocardial tissue is improved in group IPO and group P50 as compared with that in group Con,while group M + IPO is more serious than group IPO,group M + P50 is more serious group P50.The mitochondrial Flameng score is higher among each group as compared with group N (P ＜ 0.05),the score is lower in group IPO and group P50 as compared with group Con and corresponding nonblocking group (M + IPO,M + P50,P ＜0.05).The mRNA and the protein expressions of HO-1,NQO1,SOD1 and Nrf2 among each group are lower as compared with group N(P ＜0.05).Compared with those in group Con,the mRNA and the protein expressions in group IPO and group P50 are obviously increased(P ＜ 0.05),group IPO and group P50 are higher than those in group adding active oxygen scavenger(MPG) (P ＜ 0.05).Conclusion Ischemic postconditioning and pinacidil postconditioning have protective effect of myocardial tissue from ischemia reperfusion injury,while improve the cardiac function index.The cardiac protective effect of Ischemic and Pinacidil postconditioning methods may be involved the ROS in early reperfusion,which activate the Nrf2-ARE pathway,and up-regulate the expression downstream antioxidant protein and phase Ⅱ detoxifying enzyme,ultimately improve the cardiac function index during the reperfusion period.

ObjectiveTo investigate the effect ofpinacidil hyperpolarizaed arrest on p38 mitogen-activited protein kinase (p38MAPK) during myocardial ischemia-reperfusion (I/R) in isolated rat hearts.MethodsFortyeight male SD rats weighting 250-300 g were randomly divided into 6 groups( n =8 each): natural arrest group (group A) ; St.Thomas group (group B) ; pinacidil hyperpolarization arrest group (group C) ;5-hydroxydecanoate (5-HD) group (group D);HMR-1098 group(group E) and 5-HD + HMR-1098 group(group F).Langendorff reperfusion model was established and K-H solution was retrogradely perfused for 15 min.In group A the hearts were arrested naturally afar perfusion was stopped; in group B St.Thomas solution was perfused; in group C pinacidil hyperpolarization solution was perfused; in the other three groups,K-H solution was perfused to isolated rat hearts for 10 min followed by 5 min 5-HD (group D) or HMR-1098(group E) or 5-HD and HMR-1098(group F) perfusion,then hyperpolarization arrest solution was given in each group.Each hearts suffered 60 min ischemia after arrest followed by 30 min K-H solution reperfusion.Coronary flow(CF),HR,left ventricular developed pressure( LVDP),left ventricular systolic pressure(LVSP) and the maximum rate of pressure rise (dp/dtmax) were measured at the end of 15 min K-H solution perfusion and at 20 min of reperfusion.Myocardial phosphatic and nonphosphatic p38MAPK expression was determined by Western blot at 30 min of reperfusion.ResultsCompared with group C,CF,HR,LVDP,LVSP and dp/dtmax were significantly decreased at 20 min of reperfusion and phosphatic p38MAPK expression was down-regulated,non-phosphatic p38MAPK expression was up-regulated at 30 min of reperfusion in groups A,B,D,E and F (P ＜ 0.05).Compared with group E,CF,HR,LVDP,LVSP and dp/ dtmax were significantly decreased at 20 min of reperfusion and phosphatic p38MAPK expression was down-regulated,non-phosphatic p38MAPK expression was up-regulated at 30 min of reperfusion in groups D and F ( P ＜0.05).ConclusionHyperpolarized arrest induced by pinacidil can improve cardiac function following myocardial I/R injury by up-regulating phosphatic p38MAPK expression,down-regulating non-phosphatic p38 MAPK expression and mitochondrial potassium channel is more important than membranous one during the regulation of phosphatic p38MAPK expression.

Objective To analyze the dose‐effects of different doses of dexmedetomidine for sedation and sleep of the patients under epidural nerve block anesthesia .Methods A total of 82 patients undergoing elective lower limbs surgery(ASA grade Ⅰ - Ⅱ) were randomly divided into 4 different doses of dexmedetomidine groups(group 1 ,n=19 ;group 2 ,n=22 ;group 3 ,n=20;group 4 , n=21) ,under continuous epidural nerve block ,the loading dose of dexmedetomidine 0 .7 ,0 .8 ,0 .9 ,1 .0μg · kg -1 · h-1 was intrave‐nously pumped for 30 min ,then pumped at a rate of 0 .7 ,0 .8 ,0 .9 ,1 .0 μg · kg -1 · h-1 in the group 1 ,2 ,3 and 4 respectively .If any patient in 4 groups fell asleep at less than half an hour ,the loading dose was stopped and the continuous dose was changed to pump , the drug administration was discontinued at wound dressing after operation .Whether the patient falling asleep was recorded ,and the mean arterial blood pressure ,heart rate at 4 time points of 10 ,30 ,60 ,90 min after infusion of dexmedetomidine were also recorded . The 50% effective dose(ED50 ) ,ED95 and 95% confidence interval(CI) were calculated by using the Probit method .Results ED50 and ED95 of dexmedetomidine were 0 .65μg/kg(95% CI:0 .36-0 .73μg/kg)and 1 .00 μg /kg(95% CI:0 .90-1 .74μg/kg) ,which could decrease the heart rate and increase the arterial blood pressure .Conclusion Although dexmedetomidine can decrease the heart rate and increase the arterial blood pressure ,but the patients quietly fall asleep without discomfort and pain occurrence by the intra‐venous administration under continuous epidural nerve block .

Objective To investigate the role of reactive oxygen species ( ROS ) in emulsified isoflurane postconditioning?induced promotion of nuclear factor?E2 related factor 2 ( Nrf2 )∕antioxidant re?sponse element ( ARE) signaling pathway activation in rats with myocardial injury. Methods Healthy male Sprague?Dawley rats, aged 16-20 weeks, weighing 250-300 g, were heparinized and anesthetized. Their hearts were excised and perfused in a Langendorff apparatus with K?H solution. Eighty isolated rat hearts were divided into 5 groups ( n=16 each) using a random number table: control group ( group C);ischemia?reperfusion (I∕R) group; emulsified isoflurane postconditioning group (group EIP); fat emulsion postconditioning group (group FEP); N?(2?mercaptopropionyl)?glycine (a ROS scavenger) plus emulsi?fied isoflurane postconditioning group ( group N+EIP ) . Group C was perfused with K?H solution for 100 min, and the other 4 groups were subjected to 40 min of ischemia followed by 60 min of reperfusion. EIP and FEP groups were perfused for 2 min with K?H solution containing 1?68 mmol∕L emulsified isoflurane and 712 mg∕L intralipid, respectively, starting from the onset of reperfusion, and then continuously per?fused with K?H solution for 58 min. Group N+EIP was perfused for 3 min with K?H solution containing N?(2?mercaptopropionyl)?glycine 2 mmol∕L, and then emulsified isoflurane postconditioning was performed. Heart rate ( HR ) , left ventricular developed pressure ( LVDP ) , left ventricular end?diastolic pressure (LVEDP) and the maximum rate of increase of left ventricular pressure (+dp∕dtmax) were recorded at the end of equilibration and of reperfusion. At 5 min of reperfusion and the end of reperfusion, myocardial specimens were obtained for determination of ROS content. At the end of reperfusion, myocardial specimens were obtained for examination of the ultrastructure of myocardial cells and for determination of the expression of Nrf2, heme oxygenase?1 ( HO?1 ) , quinone oxidoreductase 1 ( NQO1 ) and superoxide dismutase?1 ( SOD?1) protein and mRNA in myocardial tissues. Mitochondrial injury scores ( Flameng scores) were e?valuated. Results Compared with group C, HR, +dp∕dtmax and LVDP were significantly decreased, LV?EDP , mitochondrial Flameng scores and ROS contents were increased, and the expression of Nrf2, HO?1, NQO1 and SOD?1 protein and mRNA was down?regulated at the end of reperfusion in I∕R group ( P<0?05) . Compared with group I∕R, HR, +dp∕dtmax and LVDP were significantly increased, LVEDP, mitochondrial Flameng scores and ROS contents were decreased, and the expression of Nrf2, HO?1, NQO1 and SOD?1 protein and mRNA was up?regulated at the end of reperfusion in group EIP ( P<0?05) , and no significant changes in the parameters mentioned above were found in group N+EIP ( P>0?05) . Compared with group EIP, HR, +dp∕dtmax and LVDP were significantly decreased, LVEDP and mitochondrial Flameng scores were increased, ROS contents at 5 min of reperfusion were decreased, ROS contents at the end of reperfu?sion were increased, and the expression of Nrf2, HO?1, NQO1 and SOD?1 protein and mRNA was down?regulated in group N+EIP ( P<0?05) . The degree of myocardial injury was reduced in group EIP as com?pared with group I∕R. There was no significant difference in the degree of myocardial injury between group N+EIP and group I∕R. Conclusion The mechanism by which emulsified isoflurane postconditioning pro?motes Nrf2?ARE signaling pathway activation is totally related to ROS in rats with myocardial injury.

To investigate the relationship of the variation of virulence and the external capsid proteins of the pandemic duck hepatitis A virus type 1(DHAV-1) isolates,the virulence,cross neutralization assays and the complete sequence of the virion protein 1(VP1) gene of nine virulent DHAV-1 strains,which were isolated from infected ducklings with clinical symptoms in Shandong province of China in 2007-2008,were tested.The fifth generation duck embryo allantoic liquids of the 9 isolates were tested on 12-day-old duck embryos and on 7-day-old ducklings for the median embryonal lethal doses(ELD50s) and the median lethal doses(LD50s),respectively.The results showed that the ELD5s of embryonic duck eggs of the 9 DHAV-1 isolates were between 1.9 × 106/mL to 1.44 × 107/mL,while the LD50s were 2.39 × 105/mL to 6.15 × 106/mL.Cross-neutralization tests revealed that the 9 DHAV-1 isolates were completely neutralized by the standard serum and the hyperimmune sera against the 9 DHAV-1 isolates,respectively.Compared with other virulent,moderate virulent,attenuated vaccine and mild strains,the VP1 genes of the 9 strains shared 89.8％-99.7％ similarity at the nucleotide level and 92.4％-99.6％ at amino acid level with other DHAV-1 strains.There were three hypervariable regions at the C-terminus(as 158-160,180-193 and 205-219) and other variable points in VPI protein,but which didn't cause virulence of DHAV-1 change.

AIM: To investigate the effects of the selective mitochondrial ATP-sensitive K+ channels opener diazoxide on mitochondrial respiratory function and enzyme activity in isolated rat myocardium under ischemia/reperfusion.METHODS: Observation was made on rat hearts perfused with Langendorff apparatus.72 Sprague-Dawley(SD) rats were randomly divided into 4 groups: normal group(NOR),ischemia reperfusion(IR),diazoxide group(DIA) and 5-hydroxydecanoate(5-HD) antagonized diazoxide group(5HD-DIA).Hearts isolated from SD rats were mounted on a Langendorff apparatus and started with a 20 min perfusion for equilibration.NOR went on perfusion for another 100 min after equilibration.IR underwent 40 min global ischemia and followed by 30 min reperfusion after 30 min stabilization.DIA was administered with K-H solution containing diazoxide at concentration of 50 ?mol/L for 10 min before ischemia and reperfusion.5HD-DIA was infused with 100 ?mol/L 5-HD(a specific mitochondrial ATP sensitive K+ channel blocker) and the same procedure was carried out as DIA group.Hearts were taken down to extract mitochondrial at the end-equation,before ischemia and at the end-reperfusion for determination of mitochondrial respiratory function and the enzyme activity of mitochondria.RESULTS: At the end of reperfusion,mitochondrial respiratory function(mitochondrial respiratory control rate,P/O ratio and state 3 respiration) and mitochondrial enzyme activity(NADH oxidase,succinate oxidase and cytochrome C oxidase) in DIA group were better than those in IR group and 5HD-DIA group(P0.05).CONCLUSION: Preconditioning with mitochondrial ATP sensitive potassium channel opener,diazoxide,protects rat heart mitochondria against ischemia-reperfusion injury.The mechanisms are involved in the safeguarding of respiratory function and activity of enzymes of respiratory chain.

OBJECTIVETo observe the effect difference between drug-spreading moxibustion and the oral administration of meloxicam for primary dysmenorrheal with cold stagnation and to explore its mechanism.

METHODSA total of 101 patients with primary dysmenorrheal were randomly assigned into a drug-moxibustion group(52 cases) and a western medication group(49 cases). Drug-spreading moxibustion was used on the lumbosacral acupoints area and then around lower abdominal five days before menstruation until the 3rd day of menstruation,once three days,while western medicine meloxicam was prescribed one day before menstruation,7.5 mg at a time,once a day and continuously for three days. The clinical effects after one course,namely three menstrual cycles,were compared between the two groups. Meanwhile,the resistance index(RI) and the pulsatility index (PI) of uterine artery and arcus arteriarum were examined through color Doppler ultrasound before and after treatment.

RESULTSAfter one-course treatment,the effective rate was 92.3%(48/52) in the drug-spreading moxibustion group,which was better than 67.3%(33/49) in the western medication group(<0.05). Also,all the RI and PI in the drug-spreading moxibustion group were obviously decreased than those before treatment(all<0.05),and the ones were superior to those of the western medication group(all<0.05),which showed no apparent decrease after treatment(all>0.05).

CONCLUSIONSDrug-spreading moxibustion can improve the symptoms of primary dysmenorrheal with cold-damp stagnation,and the effect is better than that of meloxicam. The mechanism may be related to improve the blood supply to the uterus.